mohloli o moholo
Setsi sa Wagoner sa Joala le lemalla ntho e Patlisiso, Setsi sa Neuroscience, Univesithi ea Texas ho Austin, Austin, TX, 78712, USA. [imeile e sirelelitsoe].
inahaneloang
NAKO E MONG:
LITLHAHISO:
Ho se khone ho fokotsa kapa ho laola tšebeliso ea joala ke letšoao le hlahelletseng bakeng sa mathata a tšebeliso ea joala. Patlisiso mabapi le boits'oaro bo botle ba boitsoaro le liphatsa tsa lefutso tse bakiloeng ke phetoho ea joala e tla ntšetsa pele tsebo ea rona ka mathata a tšebeliso ea joala. Boitšoaro bo fapaneng ba taolo ea joala bo ile ba bonoa nakong e fetileng ha bo bapisa mefuta e 'meli ea poone e nyalisitsoeng ea F1: C57BL / 6J x NZB / B1NJ (BxN) bonts'a khetho e fokotsoang ea joala kamora boiphihlelo ka ho nooa haholo le linako tsa ho itlopa joala ha C57BL / 6J x FVB / NJ (BxF) bontša likhetho tsa joala tse tsoakiloeng. Tsena tsa phenotypes lia khahla hobane li-hybrids tsena li bonts'a ho ba teng ha liphatsa tsa lefutso (BxN) le boteng bo feteletseng (BxF) ka ho kenella hoa ethanol ka tsela e itšetlehileng ka boiphihlelo.
Ka ho khetheha, BxF e bonts'a khetho e tsoakiloeng ea joala le ponts'o ea BxN e fokolitse khetho ea joala kamora boiphihlelo ka lintlha tse phahameng tsa ethanol; Leha ho le joalo, boiphihlelo bo nang le ethanol e tlase bo etsa hore batho ba se ke ba rata joala haholo ka linako tse peli.
Phuputsong e teng hona joale, re ile ra leka khopolo ea hore lipono tsena li emeloa ke tlhahiso e fapaneng ea sepheo sa transcript, ΔFosB, moputsong, ho hlohlona le likarolo tse amanang le khatello ea kelello.
LIKOTSO:
Liphetoho ho li-neuronal tsa polasetiki (joalo ka ha li lekantsoe ke maemo a ΔFosB) li ne li its'epahalla boiphihlelo, hammoho le sebaka sa boko le genotype, ho ts'ehetsa ka ho fetelletseng hore ts'ebetso ea methapo ea methapo ea kutlo.
Pontšo ea litoeba tsa BxN e fokotsitseng khetho ea joala e ne e na le maemo a tlase a ΔFosB mokokotlong oa Edinger-Westphal ho feta litoeba tse tsoetseng pele ho rata joala, mme e ile ea eketsa maemo a ΔFosB bohareng ba medy amygdala ha e bapisoa le litoeba tsa taolo.
Litoeba tsa BxN tse bonts'a khethollo ea joala e tsoetseng pele li bonts'a litekanyetso tse phahameng tsa ΔFosB sebakeng se katamelaneng sa moea, Edinger-Westphal nucleus, le amygdala (karohano e bohareng le ea morao-rao).
Ho feta moo, ka Maemo a litoeba tsa BxN ΔFosB mokokotlong oa Edinger-Westphal le libaka tse arohaneng tsa likarolo tse kenang hantle li hokahane hantle le khethollo le takatso ea ethanol. Ho ekelletsa moo, tlhahlobo ea likamano tsa bohanyetsi e ile ea senola hore litoeba tse ngata tsa ethanol-naïve tse nang le maemo a tlase a ΔFosB li ka har'a sehlopha, athe litoeba tse ngata li bonts'a khetho ea joala e nang le maemo a phahameng a ΔFosB ka sehlohlolong hammoho.
CONCLUSIONS:
Ka ho bapisa le ho bapisa phenotypes tse peli tsa joala, boithuto bona bo bonts'a hore lipakana tse amanang le moputso le khatello ea maikutlo (ho kenyeletsa le konopo ea Edinger-Westphal, sebaka sa patralal tegmental, amygdala) li ba le polasetiki ea bohlokoa e bonahatsang le litakatso tsa joala tse fokotsoang.
Background
Ho na le mabaka a tsebahalang a ho hlaseloa habonolo, tikoloho le liphatsa tsa lefutso, tse amanang le tšebeliso e mpe ea joala le bokhoba ba tahi. Bokhoni ba ho noella joala bo bongata ntle le phello e nyane ho motho ka mong ke lets'oao la mantlha ho lino tse tahang, ho bonts'a hore tekanyo e tlase ea karabelo ho joala ke sesosa se seholo sa tlhabollo ea joala.1,2]. Ho hlalosa mabaka a amanang le ts'ebetso ea joala a tla re thusa ho utloisisa ts'ebeliso e mpe ea joala, 'me ke leano le sebetsang la ntlafatso ea kalafo e ntlafalitsoeng bakeng sa batho ba nang le bothata ba tšebeliso ea joala. Ts'ebeliso ea mefuta ea litoeba ho etsisa lefu la batho e bile sesebelisoa se matla tsoelo-peleng ea ho utloisisa lefu lena le ho ntlafatsa kalafo. Hona le mehlala e mengata ea ho ithuta ka tlhekefetso ea joala le joala, leha ho le joalo ha ho na mohlala oa botahoa ka botlalo. Boima boo panya e tla itlhomella ka boeona ho fana ka tharollo ea ethanol tlasa maemo a ts'oanang a tikoloho e itšetlehile haholo ka semelo sa eona sa lefutso [3].
Haufinyane tjena re fumane hore C57BL / 6JxFVB / NJ (BxF) le FVB / NJxC57BL / 6J (FVBxB6) F1 poone e nyalisitsoeng e sa ipehele maemo a sa tloaelehang a joala nakong ea litlhahlobo tsa botlolo tse peli tsa botšehali / golf XNUM / XN le banna 20-35 g / kg / letsatsi, ho ipapisitse le mahloriso le paradigm) [4]. Moetso ona o mocha oa liphatsa tsa lefutso o na le molemo o moholo ha o bapisoa le mefuta e teng ea tlhaho e kentsoeng, ho kenyelletsa le bopaki ba mofuta o mongata haholo oa tšebeliso ea mali le ho nooa ho isa tekanyong e phahameng ea joala ba mali [4]. Ntle le moo, tšebeliso e phahameng ea ethanol e bonts'itsoeng ke litoeba tsa BxF e bonoa litsong tse ling tse peli tsa ho nwa tsa ethanol (ho nwa lefifing le kamohelo ea ethanol nakong ea phihlello ea metsi e reriloeng)4]. Joale re bone boits'oaro bo ikhethileng ba taolo ea lithethefatsi ha re bapisa mefuta e 'meli ea li-hyster tsa F1: X57BL / 6J x NZB / B1NJ (BxN) li bonts'itse khetho ea lithethefatsi kamora boiphihlelo bo tebileng ba joala le linako tsa ho itlopa joala le BxF li bontša ts'ebetso ea tahi e tsoetseng pele.5]. Re sebelisa liteko tse ngata tsa boitšoaro, re bonts'a hore BxN e tebile ho feta litoeba tsa BxF ho batho ba iphapanyetsang maikutlo a ho tsilatsila, empa e seng moputso, litlamorao tsa ethanol [6].
Patlisiso ea mantlha mabapi le boits'oaro bo sa tloaelehang ba boitšoaro le liphatsa tsa lefutso tse noang joala haholo le liphetoho tse bakiloeng ke ho nooa li tla ntšetsa pele tsebo ea rona mabapi le tšebeliso e mpe ea tahi le joala. Mokhoa o fokotsoang oa khetho ea joala o khahlisa hobane qalong litoeba tsa BxN li bontša khetho e phahameng bakeng sa tharollo ea ethanol. Leha karolo e susumetsang ea ho fokotsa ts'ebeliso ea joala kamora boiphihlelo ka tekanyo e phahameng ea ethanol le ho itlopa joala e sa tsejoe, litoeba tsa BxN li ka tšoantšoa le ba noang joala haholo ka hore ba ntse ba sebelisa tharollo ea ethanol empa ka tekanyo e fokotsehileng, mohlomong ka lebaka la boiphihlelo.
Mohlala oa ho rata joala o tsoetseng pele o boetse oa khahlisa, hobane litoeba tsa BxF ka mokhoa o ts'oanang li sebelisa maemo a phahameng haholo a ethanol ho sa tsotelehe boiphihlelo bo fetileng. Takatso ea joala e tsitsitseng le e fokotsoang e ka amana le phello ea tlhekefetso ea joala, ketsahalo eo ho eona liphoofolo li bonts'ang haholo ts'ebetso ea ts'ebeliso ea joala kamora nako ea ts'ebetso e qobelloang.e [7]. Tšusumetso ea tlhekefetso ea joala ke ntho e bohlokoa molemong oa ho ithuta ho eketsa ts'ebeliso ea lino tse tahang. Leha kemiso ea teko e tsejoang ho susumetsa ts'ebetso ea tlhabollo ea joala e fapane haholo le kemiso e sebelisitsoeng mona, ho bapisa khetho e tsitsitseng le e fokotsoang ea joala le ts'ebeliso ea lino tse tahang e amana le boits'oaro bo fapaneng ba boits'oaro bo hlalositsoeng mona ketsahalong ea bohlokoa mehlaleng ea lipatlisiso tsa joala. Khethollo ea joala e fokotsehileng e ka ba karolo ea phello ea tlhekefetso ea joala mme khetho ea joala e tsoakoang e ka hlalosoa e le ho se be teng ha phello ea tlhekefetso ea joala. Ts'ebeliso ea mefuta e fapaneng ea liphoofolo tsa liphatsa tsa lefutso, joalo ka BxF le BxN, e thusa haholo nts'etsopele ea masimo kaha ho nahanoa hore mathata a tšebeliso ea joala a hlaha likhoebisanong tse rarahaneng lipakeng tsa liphatsa tsa lefutso le tikoloho. Ho tsebahatsoa ha mofuta oa liphatsa tsa pele tsa lefutso bakeng sa mefuta ena e nyalisitsoeng ho fana ka leseli mabapi le ho potoloha hoa boko ho bohlokoa bakeng sa thepa e atlehang le e nyenyefatsang ea ethanol.
Ethanol le li-neurocircuits tse ling tse amanang le lithethefatsi li ithutile mefuteng e ikhethang ea litoeba li sebelisa matšoao a limolek'hule a polasetiki ea neuronal le / kapa ketso [8-15]. Ethanol e ikemetseng le e sebelisang liteko ha e hlahise limmapa tse tšoanang tsa metabolic ea boko, e fana ka maikutlo a hore potoloho e itseng e tlatselletsa litlamorao tsa ethanol [8,9].
Karolo e 'ngoe ea bohlokoa, e sa ntseng e lokela ho hlahlojoa haholo liphuputsong tsa joala, ke tlhahlobo ea boitšoaro bo ts'oaroang le bo fokotsehileng ba boits'oaro ba joala le ho tsebahatsa mekhahlelo ea methapo e sebetsanang nakong ea boits'oaro bona. Morero oa teko ena e ne e le ho supa libaka tsa boko tse amanang le tšebeliso e mpe ea joala le ho fokotsa. Hobane taolo e sa foleng ea tahi (hammoho le lithethefatsi tse ling tsa tlhekefetso) e bonts'itsoe ho baka likarohano tsa tikoloho ea boko maemong a ΔFosB, re lekile maikutlo a hore mehopolo ena ea boitšoaro e emeloa ke tlhahiso e fapaneng ea tšibollo e sa amaneng, ΔFosB, libakeng tsa boko tse tsejoang ho ba le seabo ho moputso, ho ferekana le khatello ea maikutlo [10].
Lisosa tse sa feleng tse bakang karohano ea tikoloho maemong a ΔFosB li kenyelletsa lithethefatsi tsa tlhekefetso (joala, cocaine, amphetamine, nicotine, morphine, le antipsychotic), khatello ea maikutlo (khatello ea maikutlo), khatello ea maoto e sa lebelloang, ho tsitsipanoa ka motlakase), le mokokotlo o mamellehang oa ho matha [11]. Joaloka mokena-lipakeng oa liphetoho tsa nako e telele bokong, ho khetholla phapang e atileng ea FosB (FosB kapa ΔFosB) karabong ea kalafo ea ethanol e sa foleng ke phapang ea bohlokoa.
Ho na le lithuto tse 'maloa tse lekantseng FosB le ΔFosB kamora ts'usumetso e sa foleng eo ho sa netefatsoang hore ΔFosB e ne e le isoform e hlaheletseng (joalo ka e hlalositsoeng ka tlase). Leha ho le joalo, ho na le bopaki bo matla ba hore ΔFosB, eseng FosB, ke isoform e hlahelletseng ka mor'a ts'usumetso e sa foleng [10-12]. Phuputso e entsoeng ke Ryabinin le Wang (1998) e fumane hore ka joala bo fokolang bo khethang litoeba tsa DBA / 2J, matsatsi a mane a ente ea enteol a phetseng esale pele a fella ka keketseho e matla ponts'o ea FosB libakeng tse latelang tsa boko: anterior cortical amygdaloid nucleus, lateral septum ventrale, amygdala central , lateral amygdala, lateral hypothalamus, nucleus e bokellang likhetla, mookotaba oa stria terminalis, le paraventricular nucleus ea thalamus [13]. Liphetho tsa bona li bontša ts'ebetso ea methapo ea kutlo e amanang le ethanol. Polelo ea FosB e boetse e lekantsoe ho joala bo phahameng ba khetha C57BL / 6J mouse nakong ea ho fumana le ho boloka tsamaiso ea ethanol tlasa maemo a fokolang a phihlello. Ha ho na liphetoho lipapatsong tsa FosB nakong ea ho iphumanela ho itaola [14]. Leha ho le joalo, kamora libeke tse peli tsa taolo e lekanyelitsoeng ea ethanol, litekanyetso tsa FosB li ile tsa eketseha mokokotlong oa methapo ea methapo ea amygdala le Edinger-Westphal nucleus [15]. Ka kakaretso, litlaleho li supa libaka tse ncha tse sebetsanang le boitšoaro ba ethanol, hammoho le ho kenya letsoho tseleng ea mesocorticolimbic tseleng le amygdala e atolositsoeng [16]. Leha ho le joalo, ho bohlokoa ho hlokomela hore liphetoho maemong a ΔFosB li its'etleha ka tsela ea tsamaiso ea ethanol, tekanyetso le bolelele ba nako bo pepesitsoeng kalafo kapa kemiso [13-15].
Mefuta ea litoeba e sebelisitsoeng thutong ena e fana ka mefuta e khahlisang ea ho bapisa khetho e tsitsitseng ea joala le mekhoa e ikarabellang bakeng sa likarabo tsena tse fapaneng tsa joala. Boithuto bona bo bonts'a hore litoeba tse bonts'ang litheko tse fokotsoang tsa joala le tsona li bonts'a polokeho ea bohlokoa lipotolohong tse amanang le moputso le khatello ea maikutlo (ho kenyelletsa le hlooho ea Edinger-Westphal, sebaka sa sephetho sa moea, amygdala, li-bokellus tsa nucleus le cingex cortex).
Results
Litlamorao tsa ho tsepamisa mohopolo oa joala le linako tsa ho itlopa joala ho ho itaola ho litoeba tsa BxF le BxN
Ho bontša hore maemo a fapaneng a ethanol le / kapa linako tsa ho itima lijo li fetotse tšebeliso ea 'nete ea ethanol, re thehile lihlahlobo tse' ne (lihlopha) ho lekanya tšebeliso ea ethanol (Setšoantšo. (Figure1a, b).1a, b). Ho bile le lihlopha tse 'ne tsa liteko bakeng sa mofuta o mong le o mong o nyalisitsoeng: Likhakanyo tse Phahameng, Likhakanyo tse Phahameng ka Nako ea Thibelo ea Bohlola, Litekanyetso tse tlase, le Likhakanyo tse tlase ka linako tsa Abstinence. Lintlha tse felletseng bakeng sa khetho ea ethanol (Setšoantšo (Figure2)2) le ts'ebeliso (Setšoantšo (Figure3)3) ya data (bakeng sa lihlopha tsohle le genotypes ka bobeli) e hlahisoa bakeng sa ho buuoa. Ho theha le ho hlakisa boits'oaro ba boitšoaro bo ts'oaroang le bo fokotsoang ba khetho ea joala, khetho ea 9% ea ethanol le datha ea ts'ebeliso e hlahisoa ho Litšoantšo LITLHOSXX4 'me and5.5. Ts'ebetso tsena tsa boitshwaro li ipapisitse le papiso ea 9% ethanol le tšebeliso ho tsoa ho tlhahiso ea pele, ea bobeli, ea boraro le ea bone ho lihlopha tse phahameng tsa Concentrations le matsatsi a liteko a tsamaeang le liteko bakeng sa lihlopha tsa Low Concerrations. Mekhoa e 'meli ea ANOVA (genotype x nako) ea khetho ea 9% ea ethanol le tšebeliso e entsoe. Bakeng sa sehlopha sa "High Concentrations", khetho ea ethanol (Setšoantšo (Figure4a)4a) le tšebeliso (Setšoantšo (Figure5a)5a) li ne li le kholo bakeng sa BxF ho feta BxN, 'me BxF e bonts'a khetho e tsoelang pele ea joala le ts'ebeliso ha BxN e bonts'a ho fokotsa khetho ea joala le tšebeliso ea eona (ETHANOL PREFERENCE - tšebelisano F (3,54) = 4.83, P <0. 01, genotype F (1,54, 24.10) = 0.001, P <3,54, nako F (9.92) = 0.0001, P <1,54; ETHANOL CONSUMPTION - tšebelisano N / S, genotype F (50.73) = 0.0001, P <3,54, nako F (11.68, EA-0.0001) = XNUMX, P <XNUMX). Bakeng sa sehlopha sa Matšoenyeho a Phahameng se nang le boithaopo, khetho ea ethanol (Setšoantšo (Figure4b)4b) le ts'ebeliso (Setšoantšo (Figure5b)5b) li ne li le kholo bakeng sa BxF ho feta BxN, 'me BxF e bonts'a khetho ea joala e tsoelang pele le ts'ebeliso ha BxN e bonts'a ho fokotsa khetho ea joala le tšebeliso ea eona (ETHANOL PREFERENCE - tšebelisano F (3,132) = 15.89, P <0.0001, genotype F (1,132) = 250.43, P <0.0001, nako F (3,132) = 27.48, P <0.0001; ETHANOL CONSUMPTION - tšebelisano F (3,132) = 11.35, P <0.0001, genotype F (1,132) = 510.88, P <0.0001, nako F (3,132) = 22.42, EA-P <0.0001) Bakeng sa sehlopha sa maemo a tlase, khetho ea ethanol (Setšoantšo (Figure4c)4c) le ts'ebeliso (Setšoantšo (Figure5c)5c) li ne li le kholo bakeng sa BxF ho feta BxN, 'me poone e nyalisitsoeng ka bobeli e bontšitse khetho e tsoetseng pele ea joala le ts'ebeliso (ETHANOL PREFERENCE - tšebelisano N / S, genotype F (1,54) = 12.2, P <0.01, nako N / S; ETHANOL CONSUMPTION - tšebelisano N / S, genotype F (1,54) = 74.83, P <0.0001, nako N / S). Bakeng sa sehlopha sa maemo a tlase se nang le boithaopo, khetho ea ethanol (Setšoantšo (Figure4d)4d) le ts'ebeliso (Setšoantšo (Figure5d)5d) li ne li le kholo bakeng sa BxF ho feta BxN, 'me poone e nyalisitsoeng ka bobeli e bonts'itse phokotso e itekanetseng ea joala le tšebeliso ea eona (ETHANOL PREFERENCE - tšebelisano N / S, genotype F (1,132) = 166.58, P <0.0001, nako N / S; ETHANOL CONSUMPTION - tšebelisano F (3,132) = 3.61, P <0.05, genotype F (1,132) = 480.64, P <0.0001, nako F (3,132) = 7.87, P <0.0001). Ka bokhutšoanyane, lihlopheng tsa Matšoenyeho a Phahameng (ntle le ho ithiba), BxF e bonts'itse khetho e tsoelang pele ea joala ha BxN e bonts'a ho fokotsa joala le lihlopheng tsa Low Concentrations (ntle le ho ithiba), ka bobeli BxF le B6xN li bontšitse khetho e tsoelang pele ea joala. Kaha li-phenotypes tsa thahasello li tšoaroa hantle ka lihlopha ntle le ho ithiba, ke tsona tsepamiso ea maikutlo ho karolo e setseng ea thuto.
Mehato ea ΔFosB
Quanifying le quFosB li sebelisitsoe ho tsebahatsa ts'ebetso ea methapo e mengata nakong ea ts'ebetso e tsoakiloeng le e fokotsoang. Ho ne ho na le lihlopha tse tharo tsa liteko bakeng sa mofuta o mong le o mong o nyalisitsoeng: Tabatabelo e Phahameng, Lits'ireletso tse tlase, le Metsi (Taolo). Lintlha tsa ΔFosB li hlahisoa e le liperesente tse ΔFosB positive neurons [(# of ΔFosB positive neurons) / (# of ΔFosB positive neurons + # of Nissl positive neurons)] (Lethathamo (Letlapa1).1). Mosebetsi oa nakong e fetileng o bontšitse hore boiphihlelo ba ethanol bo ka tsitsisa neurodegeneration [17]. Ka hona, re fuputse lipalo tsa li-neuronal thutong ena mme ha re tlalehe phapang ea bohlokoa e thehiloeng ho genotype kapa sehlopha bakeng sa libaka tsa boko tse hlalositsoeng thutong ena. Litlhatlhobo tse latelang tse tharo tsa data ea ΔFosB li entsoe: 1) mekhoa e meraro ea ANOVA (genotype x group x brain mkoa), 2) tsela tse peli ANOVA (brain area x group) bakeng sa genotype e ngoe le e 'ngoe, le 3) lipalo tsa ntlafatso li entsoe ho hlophisa khokahano ea' mapa marang-rang.
Mehato e phetoang makhetlo a mararo ANOVA (genotype x group x brain region) e senotse genotype x brain region interaction [F (15,375) = 2.01, P <.05], a group x brain region interaction [F (15.375) = 1.99, P <0.01], le phello e kholo ea sebaka sa boko [F (15,375) = 43.36, P <.000]. Mehato e phetoang habeli ANOVA (sebaka sa boko x sehlopha) bakeng sa genotype e ngoe le e ngoe e bonts'itse hore ho na le tšusumetso e kholo ea sebaka le sehlopha bakeng sa BxF le BxN [BxF - F (2,374) = 11.79, P <.0001, phello e kholo ea sehlopha; F (15,374) = 25.64, P <.0001, phello e kholo ea sebaka sa boko; BxN - F (2,360) = 43.38, P <.0001, phello e kholo ea sehlopha; F (15,360) = 23.73, P <.0001, phello e ka sehloohong ea genotype]. Tlhahlobo ea Post-hoc e senotse liphapang tse tšeletseng tsa sehlopha bakeng sa BxN (Setšoantšo (Figure6a-c).6ac). Mefuta ea Percent ΔFosB e ne e phahame ho sehlopha sa Low Conrivation ho feta sehlopheng sa Metsi se La, CeC / CeL, EW, le VTA. Percent ΔFosB e ne e phahame sehlopheng se phahameng sa tlhaloganyo ho feta sehlopheng sa Metsi se CeMPV. Percent ΔFosB e ne e phahame sehlopheng sa maemo a tlase ho feta sehlopheng sa High Concentrations ho EW. Lintlha tsa ΔFosB tsa likarolo tsohle tse ling tsa boko tse boletsoeng li hlahisitsoe Lethathamong Tafole1.1. Pearson's r correlational analysis e sebelisitsoe ho fumana hore na% ea ΔFosB li-neuron tse ntle sebakeng se fanoeng sa boko se amanang le ts'ebeliso ea ethanol kapa khetho. Ts'ebeliso ea Ethanol le khetho li bonts'a kamano e ntle le% ΔFosB ho EW le VTA ea litoeba tsa BxN (ETHANOL CONSUMPTION - EW r = 0.85; VTA r = 0.85; ETHANOL PREFERENCE - EW r = 0.83, VTA r = 0.88; p <0.05 bakeng sa bohle).
Kamano e rarahaneng pakeng tsa polelo ea ΔFosB, genotype, sebaka sa boko, le ts'ebeliso ea ethanol e ile ea hlahlojoa hape ho sebelisoa tlhahlobo ea karolo ea motheo le ho kopana ha maemo. Tlhahlobo ea likarolo tsa mantlha e senotse hore boholo ba ho fapana (~ 80%) ho data bo emeloe ke likarolo tsa 5. Ho kopanyetsoa ho sa kopaneng hoa matla (ho kopantsoeng ke batho le libaka tsa boko) ho ne ho etsoa le ho laeloa ho sebelisoa karolo ea mantlha ea mantlha (Setšoantšo. (Figure7).7). Sehlopha seo ka seng se senotse mekhoa e matla, empa e sa phethahala, ea mekha ea sehlopha e thehiloeng ts'ebelisong ea ethanol, ho sa tsotelehe mofuta oa genotype. Bongata ba litoeba tsa ethanol-naïve li kopantsoe hammoho 'me li bonts'a overallFosB e fokolang ho feta se boleloang ke litoeba tse ngata tse bonts'itseng boikhethelo ba joala bo kopaneng bo bonts'a ΔFosB ka kakaretso ho feta se boleloang. Lihlopheng tsena ka bobeli e ne e le tse arohaneng haholo. Mekhahlelo e meraro e lipakeng e ne e emetse motsoako o moholo ho feta o ka tlase ho boleng ba "osFosB" le "ethanol" phenotypes.
Puisano
Boitšoaro bo fapaneng ba taolo ea joala bo ile ba bonoa ha bo bapisa mefuta e 'meli ea poone e nyalisitsoeng ea F1: BxN e bontša ho fokotseha ha lithethefatsi tsa joala kamora boiphihlelo ka botahoa bo bongata le linako tsa ho itlopa joala ha BxF e bonts'a khetho ea joala e tsoetseng pele.. Mefuta ea BxF e tsitsitseng, tšebeliso e phahameng ea joala (likhetho tsa joala tse tsoakiloeng) le mefuta ea BxN ho nooa ka tsela e itekanetseng (khetho e fokolang ea joala). Neuronal plasticity (kapa ketso, joalo ka ha e lekantsoe ke maemo a ΔFosB) e ne e fapane ho latela boiphihlelo ba ethanol, e ts'ehetsa karolo ea mantlha ea potoloho e itseng ea methapo molemong oa ho rata le ho fokotsa joala.
Bakeng sa khatello e kholo ea joala, C57BL / 6, khetho ea ethanol le ts'ebeliso li itšepa haholo ka khatello ea mantlha ea ethanol, bolelele ba ho ila, le khatello e tlase (C57BL / 6Cr kapa C57BL / 6J) [7,18]. Re fumane hore boikhethelo le tšebeliso ea ethanol tse bonoang ho litoeba tsa BxF li ne li phahame ka linako tsohle (hape li tsitsitse ho feta ho BxN) lits'ebetsong tse nne tse fapaneng tse lekiloeng. Khetho le lithethefatsi tse phahameng tsa ethanol tse itekanetseng ho BxN li ile tsa ts'oaroa feela le kemiso e le 'ngoe ea ho nwa ho sa foleng (Litekanyetso tse tlase ntle le ho itlopa joala), ha litokollo tsa ho se rate le tšebeliso li ne li bonoa le litekanyetso tse ling tsohle tsa ho noella tse sa foleng. BxN e fokolitse khetho ea joala e fana ka mofuta oa phoofolo ea mofuta oa boiphihlelo moo (boiphihlelo bo phetoang ba ethanol kamora boiphihlelo ka lintlha tse ngata tse phahameng tsa ethanol le / kapa nako e khuts'oane ea ho ila) ho fokotsa karabelo ea bona molemong oa khatello ea ethanol eo pele e neng e le eona.
Ethanol e ikemetseng le e sebelisang liteko e hlahisa limmapa tsa metabolic tsa boko tse fapaneng, e fana ka maikutlo a hore potoloho e itseng e tlasa sephetho sa matlafatso ea ethanol [8,9]. Re letile hypothesis ea hore litekanyetso tsa boitšoaro tse ts'oaroang le tse fokotsoang tsa tšebeliso ea joala li emeloa ke tlhahiso e fapaneng ea sepheo sa ho ngola se sa sebetseng, ΔFosB, libakeng tsa boko tse tsejoang ho kenya letsoho moputsong, ho hlohlolong le khatello ea maikutlo. ΔFosB ke ntho e hatisang e nang le botsitso bo ikhethang ba nako e telele ebile ha e rate ho ts'oaroa joalo ka c-Fos, empa e ipokella nakong ea kalafo e sa foleng. Keketseho ea ΔFosB e bakiloe ke ts'ebetso e eketsehileng ea neuronal mme ho nahanoa hore e bonts'a polasetiki ea nako e telele ea neuronal. Re fumane hore liperesente tsa li-neurons tse ntle tsa ΔFosB libakeng tsa boko li itšetlehile ka genotype (BxF le BxN) le sehlopha (taolo ea Metsi, Likhakanyo tse tlase, le Likhakanyo tse phahameng).
Fkapa BxN, tlhahlobo ea post-hoc e senotse hore ts'ebeliso ea ethanol ea boithaopo e hlahisitse keketseho ea ΔFosB bokong ba EW, VTA, le amygdala: e bonts'a polasetiki ea neuronal e libakeng tsa boko tse tsejoang ho nka karolo ho ethanol, moputso, le likarabo tsa khatello ea maikutlo. Litoeba tsa BxN sehlopheng sa High Concentrations (ho rata joala bo fokolisitsoeng) li fokolitse polasetiki ea neuronal ho EW, li fana ka tlhahiso ea hore li-neurons tsena li arabele tahi ea joala ka polasetiki e nang le boiphihlelo. Ho sehlopha sa Low Concentrations (ratang ho bontša joala bo tsoakiloeng), polasetiki ea neuronal ho EW e kholo ho feta lihlopheng tse phahameng tsa sehloho le metsi. Le ha e entsoe ka ho sebelisa li-paradigms tsa "ethanol" tse fapaneng le lipeo tsa liphatsa tsa lefutso, seo re se fumaneng ho EW ea litoeba tsa BxN se lumellana le lithuto tse fetileng tsa tšebeliso ea ethanol [14,15]. EW e seng preganglionic EW e sa tsoa supa e na le perioculomotor urocortin (Ucn) -container neurons [19]. Ucn1 ke cepticotropin releasing factor (CRF)-peptide e kopanyang CRF1 le CRF2 receptors. Boithuto bo fetileng bo sebelisa mekhoa ea liphatsa tsa lefutso, pharmacological, le lesion li bonts'itse hore Ucn1 e nka karolo ho laola tšebeliso ea joala [19-22]. Tmona ke tšebeliso ea lefutso e tsebahalang bakeng sa tšebeliso e phahameng ea joala ho litoeba tse lumellanang le maemo a phahameng a motheo a Ucn1 ho EW le LSi [23]. Kahoo, ho haella ha bohlokoa ba post-hoc eo re e boneng ho EW bakeng sa joala bo phahameng ba ratang le ho ja litoeba tsa BxF ho ne ho sa lebelloa. Mohlomong sena se bakoa ke liperesente tse nyane haholo tsa ΔFosB tsa sehlopha sa metsi sa BxF ha se bapisoa le sehlopha sa metsi sa BxN. Ho joalo, lipalo tsa ΔFosB ea liperesente bakeng sa litoeba tsohle tse bonts'ang khethollo ea joala (sehlopha sa BxF High Concentrations, sehlopha sa BxF Low Concentrations, le sehlopha sa BxN Low Concentrations) li ne li ts'oana.
Bakeng sa BxN, tšebeliso ea ethanol sehlopheng sa Low Concentrations e ile ea eketsa polasetiki ea neuronal ho VTA (e kholo ho feta ho Lihlopheng tse Phahameng le lihlopha tsa taolo ea Metsi). Khetho le tšebeliso ea Ethanol le tsona li ne li le kholo ho sehlopha sa Low Concentrations. Khaello ea bohlokoa ba kamora 'hoc' eo re e boneng ho VTA bakeng sa lithethefatsi tse tahang tse nang le taolo ea BxF e ne e sa lebelloa, 'me e ka ba ka lebaka la maemo a tlase a ΔFosB sehlopheng sa taolo ea metsi. Liperesente tsa ΔFosB e ne e phahame hanyane sehlopheng sa metsi a BxF ha e bapisoa le sehlopha sa metsi sa BxN, athe liperesente tsa ΔFosB li ne li batla li tšoana hantle bakeng sa litoeba tsohle tse bonts'ang khetho ea joala (sehlopha sa likhakanyo tse phahameng tsa BxF, sehlopha sa BxF Low Concentrations) . Sisteme ea VTA dopamine e bapala karolo ea bohlokoa ho hokahanya litlamorao tsa matla a ethanol mme e nka karolo mekhoeng e mengata ea ho hokahanya bohlokoa bakeng sa boitšoaro bo amanang le ethanol le boits'oaro bo amanang le moputso [24-26]. Ho feta moo, merero ea VTA e sebetsa ho li-amygdala le EW nucleus. Likhoto li bontšitsoe ho itaola ethanol ka kotloloho ho VTA [27]. Hape, ho pepeseha ha ethanol ho eketsa lebelo la ho thunya ha methapo ea methapo ea dopaminergic ho VTA [28,29]. Tekanyetso e ntseng e eketseha ea ho thunya e ka hokahana le ho kenngoa ha ΔFosB ho VTA eo re e boneng e latela taolo ea ethanol e sa fetoheng ea boithaopo ho BxN.
Ho itšetleha ka joala ho etsa hore ho be le neuroadaptations ea nako e telele, ho fella ka maemo a mabe a maikutlo mochine oa bohlokoa oa matlafatso e mpe ke corticotropin-releasing factor (CRF) ho supa kahare ho amygdala [30]. Litla-morao tsa meriana ea li-neurons ho CeA li lebisitse ho GABA, CRF, opioid, serotonin, dynorphin le li-receptor tsa norepinephrine [25,31-34]. GBA bahanyetsi, hammoho le bahanyetsi ba CRF, ba fokotsa ts'ebeliso ea ethanol [32,33,35]. Mefuta ea CeA e fokotsa ts'ebeliso e tsoelang pele ea tšebeliso ea boithaopo [36]. Seo re se fumaneng se tšehetsa karolo ea CeA ho melao ea boitšoaro ea ho tahoa. Li-neurons tsa GABAergic tse bohareng ba amygdala li theha sechaba se nang le matla a mangata se amanang le litaba tsa peptide. Li-neuron tsena tsa GABAergic li kopanya tšebetso ea CeA. Joalokaha e hlahlojoa ho [Wee le Koob (2010]), slithuto tsa kamehla li senotse karolo ea dynorphin le kappa opioid receptors ho lokiseng le ho nyoloha ha ethanol intake [37]. Haufinyane tjena, Walker et al o bontšitse hore mohanyetsi oa κ-opioid receptor, kapa-binaltorphimine, kahara amygdala ka mokhoa o ikhethileng o fokotsa taolo ea ethanol ho liphoofolo tse its'epileng [38]. Kappa opioid receptor signaling e ntse e le thahasello ea mantlha ea ho etsa lipatlisiso kopanong ea khatello ea maikutlo, moputso le boiphetetso. Ho boetse ho bontšitsoe hore tsamaiso ea khatello ea maikutlo e khothalletsoang ke khatello ea maikutlo e kopantsoe ke kappa opioid receptor signaling [39]. CeA e bohareng e ka aroloa kahare ea latero-capsular (CeL / CeC) le medial posterior ventral. Li-neurons tsa GABAergic tsa CeL / CeC li amohela li-innervation tsa dopaminergic tse tsoang VTA; Joalokaha ho hlokometsoe pejana, li-neuron tsena li kenngoa ts'ebetsong ka mor'a taolo e matla ea ethanol mme li bonts'a litoeba tse eketsehang tsa ΔFosB tse bontšang ho rata joala ho sa feleng. Hape, bona Mc [Monyaluoa (2002]) bakeng sa tlhahlobo e ntle mabapi le CeA le litlamorao tsa joala [40]. Thutong ea rona, litoeba tsa BxN tse nang le khetho ea joala e tsoetseng pele (Sehlopha sa Low Concentrations) li bontsitse polasetiki e eketsehileng ea li-neuronal ho CeC / CeL le litoeba tsa La le BxN tse nang le khetho e fokolang ea joala (sehlopha sa High Concentrations) se bonts'a polasetiki ea neuronal ho CeMPV. Liphetho tsena li fana ka maikutlo a hore boiphihlelo bo khethehileng ba ethanol bo kenyelletsa polasetiki ho GABAergic neurons ho amygdala. Ka data ena, hammoho le liphetoho tse tsamaellanang le polasetiki ea neuronal ho VTA le EW, re fana ka tlhahiso ea hore potoloho ena e na le polasetiki e tlase tlasa maemo a khethang a khethiloeng a joala.
Patlisiso e fetile e bonts'itse hore litoeba tsa C57BL / 6J li ka fihlella maemo a phahameng a joala ba mali ka libotlolo tse peli tsa khetho ea botlolo, leha ho le joalo maemo ana a joala a mali ha a ts'oaroe 'me hangata ho nooa ha ho na litekanyetso tsa khothatso ea pharmacological e boletsoeng ke Dole le Gentry (1984) [41,42]. Pontšo ea litoeba tsa BxN e bonts'itse khetho ea joala e fokotsoang ho feta e neng e ka lebelloa ho tsoa ho C57BL / 6J mouse e tloaelehileng [1]. Ka hona, leha re sa ka ra nka disampole tsa joala ba mali, ha ho na monyetla oa hore litoeba tsa BxN tse bonts'itseng khetho e batsi ea joala li fihlelle maemo a amanang le tšebeliso ea mali ka bongata, a fana ka maikutlo a hore joala bo bongata ba mali ha bo hlokehe ho kenya polasetiki libakeng tsena tsa boko. Ho bohlokoa ho hlokomela hore phello e kholo ea sehlopha le eona e teng ho BxF, leha sephetho sa morao-rao (se lokiselitsoeng bakeng sa lipapiso tse ngata) bakeng sa libaka tsa boko ba BxF se sa ka sa supa liphetoho tse kholo lipereseng tsa ΔFosB li-neurons tse ntle bakeng sa tikoloho efe kapa efe e latelang ts'ebeliso ea kamehla ea ethanol ka linako tse fapaneng.
Bakeng sa ho bona likamano tse ka bang teng lipakeng tsa li-hierarchical cluswing li entsoe. Mofuthu oa tlhahlobo e hlahisitsoeng o bonts'a mokhoa o akaretsang lipakeng tsa maemo a ΔFosB le tšebeliso ea ethanol ho sa tsotelehe mofuta oa genotype. Mefuta e phahameng ea ΔFosB e ne e amahanngoa le ho nooa haholo le litekanyetso tse tlase tsa ΔFosB li ne li amahanngoa le liphoofolo tse laolang; leha ho le joalo, matla a kamano a ne a sa lekana ho bolela esale pele li-phenotypes tse nepahetseng ho latela feela maemo a ΔFosB.
Nahanisisa
Boitšoaro bo fapaneng ba taolo ea joala bo ile ba bonoa ka mefuta e 'meli ea poone e nyalisitsoeng ea F1: BxN e bonts'a khethollo ea joala kamora boiphihlelo bo bongata ba joala ha BxF e bontša ho rata joala ho sa khaotseng. Mefuta ea BxF e tsitsitseng, tšebeliso e phahameng ea joala (likhetho tsa joala tse tsoakiloeng) le mefuta ea BxN ho nooa ka tsela e itekanetseng (khetho e fokolang ea joala). Liphetoho ho li-neuronal tsa polasetiki (joalo ka ha li lekantsoe ke maemo a ΔFosB) li ne li its'epahalla boiphihlelo, hammoho le sebaka sa boko- le genotype, li ntse li hlalosa ho potoloha ha methapo ea methapo molemong oa tšebeliso ea ethanol. Liphetho tsena li bonts'a hore phetoho ea mola o le mong oa batsoali ho litoeba tsa lebasetere e fella ka phetoho mokhoeng oa joala le liphetoho tse tšoaelitsoeng lipuong tsa polelo ea ΔFosB, e fana ka maikutlo a hore likhokahano tsa boko tse ikhethileng li etsa karolo ea litoeba tsena tse fapaneng tsa lebasetere.
mekhoa
Ethics
Boithuto bona bo ile ba etsoa ho latela likeletso tse tataisong ea Tataiso ea Tlhokomelo le Ts'ebeliso ea Liphoofolo tsa Laboraro tsa Setsi sa Naha sa Bophelo. Protocol e amohetsoe ke Komiti ea Tlhokomelo ea Liphoofolo le Ts'ebeliso ea Institutional ea Univesithi ea Texas ho Austin (AUP 2010-00028). Ho buoa hohle ho ile ha etsoa ka tlasa-sodium pentobarbital anesthesia, 'me boiteko bohle bo entsoe ho fokotsa mahlomola.
Animals
Boithuto bo entsoe ka ho sebelisa litoeba tse nyane tsa basali tsa F1 tse tsoang C57BL / 6J le litoeba tsa FVB / NJ kapa NZB / B1NJ (BxF F1 le BxN F1. C57BL / 6J, FVB / NJ, le NZB / B1NJ bareki ba ile ba rekoa ho The Jackson Laborator (Bar Haraka, ME) mme ba nyalana le 7-8 libeke. Makhoakhoa a khoesitsoeng a arotsoe ka lihlopha tse sa hlakoheng tsa mofuta o mong le o mong (BxF F1, BxN F1). Re lekile litoeba tsa basali feela ho etsa papiso le tlhaiso-leseling e neng e bokelletsoe pele [1,5,6]. Linta li ne li bolokiloe ka har'a mekotla e tloaelehileng ka lijo le metsi ad libitum. Kamore ea kolone le kamore ea ho hlahloba e ne e le lebone la 12 h: 12 h potoloho e lefifi (mabone ho 07: 00).
Teko ea likhethong tse peli tsa khetho ea botlolo
Mokhoa o mabeli oa khetho ea botlolo o sebelisitsoe ho tsebahatsa litekanyetso tsa boithatelo tsa ethanol tsa boithatelo ho litoeba tsa BxF le BxN [1,6]. F1 li-mouse tsa basali tse nyalisitsoeng (matsatsi a 63) li ne li bolokiloe ka kotlolloe mekotleng e tloaelehileng ha ba ntse ba lula beke e le 'ngoe ka libotlolong tse nang le methopo ea sipper e nang le metsi pele ho hlahisoa tharollo ea ethanol. Kamora ho lula teng, litoeba li ile tsa fumana libotlolo tse peli tse ts'oanang: e 'ngoe e na le metsi, e' ngoe e na le tharollo ea ethanol. Boemo ba Tube bo ile ba fetoloa letsatsi le letsatsi ho laola khetho ea maemo. Ho ikarabella bakeng sa ho senya le ho fetoha mouoane, boima bo tloaelehileng bo neng bo theoha ho tloha metsong ea likhaketeng tse laoloang ntle le litoeba bo ne bo tlosoa litekanyetsong tsa motho ka mong tsa ho nooa letsatsi le leng le le leng. Litsuonyana li ne li lekantsoe matsatsi ohle a 4 ho pholletsa le liteko. Ts'ebeliso eohle ea mokelikeli e ne e lekantsoe letsatsi le letsatsi ho pholletsa le liteko. Palo ea ethanol e jeoang le thatohatsi ea ethanol li baliloe ho panya e 'ngoe le e' ngoe, 'me litekanyetso tsena li lekantsoe bakeng sa karolo e ngoe le e' ngoe ea ethanol. Kameho ea ho tsepamisa mohopolo oa joala le linako tsa ho itlopa joala ho boits'ireletso ho BxF le litoeba tsa BxN li bontšitsoe ka ho qaqisa sehlopha sa liteko ka phihlello ea High Concentrations (ho phahama ho fihlella litharollo tsa ethanol tsa 3-35%, ebe ho lateloa ke 3 potoloho ea 9, 18, le 27% ethanol, e qetella ka tlhahiso ea ho qetela ea 9% ethanol) le sehlopha se seng se nang le Concentrations e tlase (ho phahamisa phihlello ea 3-9% ethanol, ka karolo e setseng ea tlhahlobo e entsoeng ka phihlello ea 9% ethanol). E 'ngoe le e' ngoe ea lihlopha tsena e ne e e-na le sehlotšoana se neng se sa sebetse ka nako e le 'ngoe ka beke. Likhoto tsa taolo li bile le maemo a tšoanang ka nako e le ngoe le litoeba tsa liteko, empa li fuoe botlolo e le 'ngoe ea metsi.
Ka kakaretso, ho ne ho e-na le lihlopha tse hlano bakeng sa lebasetere le leng le le leng: Metsi (n = 14-16), High Concentrations (n = 10), High Concentrations with Abstinence Periods (n = 20), Concentrations Low (n = 10), le Concentrations Low ka linako tsa bootsoa (n = 20). Sheba setšoantšo Figure11 bakeng sa li-schedule tsa lihlopha tse peli tse khethiloeng tsa botlolo.
ΔFosB Immunohistochemistry le quanifying
OsBFosB immunohistochemistry (IHC) e lekantsoe libakeng tsa boko ba 16 ho tloha ho litoeba tse bileng le matsatsi a 72 a phihlello e sa khaotseng ea metsi (Taolo) kapa metsi le joala [Matšoao a Holimo le Maiketsetso a tlase]. Phello ea likhakanyo tse phahameng ho khetho ea ethanol le ts'ebeliso e ne e le kholo haholo ho feta phello ea ho ithiba; ka hona, lihlopha tse bileng le linako tsa ho ithiba ha lia kenyelletsoa litekanyetsong tsa ΔFosB IHC. Ho feta moo, teko e entsoe ho feta ponahalo ea pele ea joala bo matlafalitsoeng kapa bo fokotsoang ho bonts'a boits'oaro ba phenotypes bo tsitsitse ka makhetlo a mangata a liphetoho tsa khatello ea ethanol ho lekola litlamorao tsa ts'ebeliso e sa foleng ea ethanol. Lihora tse 'ne ho isa ho tse robeli kamora ho tlosa joala ka letsatsi la 73 la teko, litoeba li ile tsa thethesoa ka matla (175 mg / kg sodium pentobarbital)' me ea sebelisoa ka mokhoa o sa sebetseng ka 20 ml ea 0.01 M phosphate buffered saline (PBS), e lateloa ke 100 ml ea 4% paraformaldehyde ho PBS. Boko bo ile ba tlosoa, ba hlophisoa kamorao ho 4% paraformaldehyde ho 4 ° C, ea kenngoa ka 3% agarose, ea aroloa (50 um, coronal) ho vibratome, ea beoa ka cryoprotectant (30% sucrose, 30% ethylene glycol le 0.1% polyvinyl pyrrolidone ho PBS) bosiu bo le bong ho 4 ° C, ebe e bolokoa ho -20 ° C ho fihlela e sebetsoa bakeng sa IHC. Likarolo tsa li-thawed li ile tsa hlatsuoa ka PBS, tsa alafshoa ka 0.3% H2O2, 'me tsa kenella hora e le' ngoe ho 3% serum e tloaelehileng ea poli ho fokotsa ho ngolisoa ho sa ikhethang. Likarolo tsa lisele li ile tsa koaheloa bosiu bo le bong ho 4 ° C ho 3% serum e tloaelehileng ea poli le anti-FosB (SC-48, 1: 5000 dilution, Santa Cruz Biotechnology, Santa Cruz, CA). Likarolo li ile tsa hlatsuoa, tsa kenella ka har'a biotinylated poli anti-mmutla Ig (1: 200 dilution, Vector Laboratories, Burlingame, CA) ka hora e le 'ngoe, ea hlatsuoa le ho kenella mohahong oa avidin-biotin (1: 200 dilution, Elite kit-Vector Laboratories) . Mosebetsi oa Peroxidase o ile oa bonoa ka karabelo ka 0.05% diaminobenzidine (e nang le 0.015% H2O2). Likarolo tsa litšiea e ne e le karolo ea Nissl e felisitsoe (a sebelisa methylene Blue / azure II). Ho ne ho behiloe li-slide bakeng sa ho bala mahlo. Li-neurons tsa ΔFosB-IR li ile tsa baloa ho ntlafatso ea 50X (oli) li sebelisa mokhoa oa ho khetholla likarolo le software ea khomphutha ea StereoInarcherator. Tlhahisoleseling ea paramente: sampore ea ho bala (50um x 50um x 10um) e ne e tšoana ka libaka tsohle tse lekanang; leha ho le joalo, boholo ba gridi bo ne bo reretsoe sebaka se seng le se seng sa boko ho netefatsa hore lipalo tse fapaneng tsa sele ea bonngoe li tla lekana 100-300 e le hore li ka fihlela katleho e lekaneng ea phapang e ka tlase ho 0.1. Lintlha li ne li baloa e le liperesente tsa ΔFosB positive nuclei (palo ea ΔFosB positive nuclei / palo ea li-neurons) sebakeng ka seng.
Antios ea FosB e sebelisitsoeng thutong ena (SC-48, Santa Cruz Biotechnology, Santa Cruz, CA) e phahamisitsoe khahlanong le sebaka se ka hare sa FosB mme e amohela FosB le ΔFosB ka bobeli. Le ha anti antianti ena e amohela bobeli ba FosB le osFosB, li-neurons tse thibelang bokhachane tse hlalositsoeng thutong ena li tla bitsoa "li-neurons tse ntle tsa ΔFosB" kaha ho bontšitsoe hore lithethefatsi tsa tlhekefetso, ho kenyelletsa joala, li susumetsa ΔFosB ka kotloloho eseng FosB. Perrotti et al. ([2008]) lekanya indFosB ho kenella (ho arabela taolo e sa fetoheng ea lithethefatsi tsa tlhekefetso, ho kenyeletsa le joala) ho sebelisa li-antibodies tse peli: e 'ngoe e amohelang FosB le ΔFosB (SC-48) le ngoe e khethang ΔFosB (ha e fumanehe khoebong) mme e fumane hore lithethefatsi tsohle ithutile, immunoreactivity e bonoang e sebelisa antio ea FosB (SC-48) e bakiloe ke ΔFosB, hobane ha ba ka ba bona li-neuron tse ling tse sa sebetseng ba sebelisa antibody khethang bakeng sa FosB ea bolelele bo felletseng [10]. Ntle le moo, ΔFosB e tsejoa ho kenyelletsa tikolohong ea boko- le mofuta o khethehileng oa sele, ka mekhoa e fapaneng ea kalafo e sa feleng le litlhahlobo tse ntle mabapi le sehlooho sena lia fumaneha [11,43,44].
Selelekela le libaka tsa sebopeho sa neuroanatomical
Il - infralimbic cortex (+ 1.70 mm); Cg1 - cingate cortex 1 (+1.1 limilimithara); Cg2 - cingate cortex 1 (+1.10 mm); Motsoako oa NAcc - nucleus accumbens core (+ 1.10 mm); Khetla ea NAcc - nucleus accumbens shell (+1.10 mm); LSi - lateral septum e bohareng (+1.10 mm); La - amygdala ea morao-rao (-1.22 mm); Bla - amygdala ea mantlha (-1.22 mm); CeC / CeL - li-capsular tse bohareng le bohareng ba lateral amygdala (-1.22 mm); CeMPV - karolo e bohareng ea posterioventral ea karolo e bohareng ea amygdala (-1.22 mm); PAG - moriri o mosoeu oa periaquaductal (-3.64 mm); EW - Edinger-Westphal khubung (-3.64 mm); VTA - sebaka sa karoloana ea sekhahla (−3.64 mm); DR - rorsal raphe (- 4.60 mm); PBN - mokokotlo oa parabrachial (-5.2 mm); NTS - nucleus tractus solitarius (-6.96 mm). Khohlo ea Mouse Stereotaxic Coordinates[45] e sebelisitsoe ho lumellanya ka nepo karolo e le 'ngoe ho isa ho e meraro bakeng sa karabelo ea karolo ka' ngoe ea boko.
Tsamaiso ea lipalo
Lintlha li tlalehiloe e le moelelo oa "SEM" ntle le moo ho boletsoeng ka tsela e 'ngoe. Lintlha li ne li ajoa ka tloaelo. Lipalopalo li entsoe ka mofuta oa Statistica 6 (StatSoft, Tulsa, OK, USA) le mofuta oa GraphPad Prism 4.00 (GraphPad Software, San Diego, CA, USA). Mehato e phetoang makhetlo a mabeli li-ANOVA li ile tsa etsetsoa tšebeliso ea ethanol le data ea khetho ho hlahloba phapang lipakeng tsa lihlopha. Li-ANOVA tse peli le tse tharo li ile tsa etsetsoa data ea ΔFosB ho lekola litšebelisano le litlamorao tse kholo bakeng sa sehlopha (Likhakanyo tse Phahameng, Tlatsetso e tlase, le Metsi), sebaka sa boko, le genotype. Khalemelo ea Bonferroni bakeng sa papiso e ngata le poso ea Bonferroni e ile ea etsoa ha ho loketse. Haholo-holo re ile ra nahana hore khatello ea maikutlo le meputso e ka be e eketsehile FosB ka litoeba tse bonts'itseng boikhethelo ba joala. Bakeng sa sefapano se seng le se seng sa lebasetere, sefapano sa Pearson se ne se sebelisetsoa ho supa boteng ba ho lumellana ha bohlokoa lipakeng tsa maemo a ΔFosB le khetho ea ethanol le tšebeliso ho litoeba tse nang le ethanol.
Ho kopanya mantsoe ka mokhoa oa Hgerarchical ho entsoe ka sepheo sa ho bona ka mahlo hore na data e hokahana joang le ho lekola hore na sehlopha sa data se kopane joang. Melao e kenelletseng ea Median e nkile sebaka sa tlhaiso-leseling ea lintlha tsa ΔFosB, tse sa feteng 15% ea data. Leha ho na le tekanyo e kholo ea ho hloka bonnete ho feta haeba melao e boletsoeng e hlile e hlokometsoe, tlhahlobo ea maemo a hloahloa ea bolaoli e hloka setho se felletseng kapa ho hlakoloa ka botlalo bakeng sa papiso e bohlale. Ho kopanya metsoako ea Hierarchical ho entsoe ka mokhoa oa Ward 'me lihlotšoana tse ileng tsa latela li laetsoe ke karolo ea pele ea tlhahlobo ea karolo ea mantlha (JMP®, Version 8, SAS Institute Inc., Cary, NC). Bakeng sa lihlopha tse nang le metsi le ethanol, tlhaiso-leseling ea ΔFosB sebakeng ka seng sa boko e ne e fetotsoe ka z-lintlha mme tlhahlobo ea likarolo tsa mantlha e entsoe ho tseba palo ea lihlopha. Lintlha li ne li kopantsoe ke likarolo tsa bokong le batho ba sebelisang tlhahlobo ea methapo ea methapo e hlokometsoeng.
Menehelo ea bangoli
ARO, YAB, RAH, TAJ ba kentse letsoho ho etseng moralo oa thuto. ARO e fumane data. ARO, IP, RDM e hlahlobile lintlha. ARO, RDM, IP, TAJ, YAB le RAH ba ne ba kentse letsoho ho ngola le ho ntlafatsa buka e ngotsoeng ka letsoho. Bangoli bohle ba ile ba bala le ho amohela buka e ngotsoeng ea ho qetela.
Liteboho
Re rata ho leboha Drs. Jody Mayfield le Colleen McClung bakeng sa lipuisano tse thusang le Marni Martinez, Jennifer Stokes, Michelle Foshat, Jose Cienfuegos, Jamie Seymour, le Darshan Pandya bakeng sa thuso ea tekheniki. Patlisiso ena e ne e tšehelitsoe ke Integrative Neuroscience Initiative on Alcoholism Consortium Grant AA13520, le National Institute on Alcohol Abuse and Alcoholism Grants AA06399-S le AA16424.
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